Aerosol compositions

ABSTRACT

The use of a medium-chain fatty acid triglyceride as the dispersant in the preparation of a medicinal aerosol composition comprising tricyclic compound (I) dispersed in a liquefied hydrofluoroalkane propellant is described.When a liquefied hydrofluoroalkane is added to a kneaded premix of the tricyclic compound (I) and a medium-chain fatty acid triglyceride, the active ingredient is evenly dispersed in the liquefied hydrofluoroalkane.Therefore, by distributing a dispenser first with the kneaded premix and, then, with a liquefied hydrofluoroalkane under cooling or elevated pressure, there can be provided a medicinal aerosol composition an improved uniformity of content of the active ingredient.

This application is a Continuation of U.S. application Ser. No.09/029,863, filed on Apr. 22, 1998, now U.S. Pat. No. 6,361,760, whichis a 371 or PCT/JP96/02670, filed Sep. 18, 1996.

TECHNICAL FIELD

This invention relates to a medicinal aerosol composition and a processfor the preparation of the same and, as such, finds application in thefield of medicine.

BACKGROUND ART

A tricyclic compound (I) and a pharmaceutically acceptable salt thereofused in the present invention have been known to possess excellentpharmacological activities such as an immunosuppressive activity and anantimicrobial activity, thereby being useful for treating and/orpreventing rejection by organ-transplantation or tissue-transplantation,graft-versus-host diseases, various autoimmune diseases and infectiousdiseases (for example, see EP-A-0184162 and WO 89/05304).

Particularly, compounds referred to as FR900506(=FK506), FR900520,FR900523 and FR900525 which belong to the tricyclic compound (I) areproduced from genus Streptomyces, in particular, Streptomycestsukubaensis No. 9993 (Depositary Authority: 1-3, Higashi 1 chome,Yatabe-machi, Tsukuba-gun, Ibaraki-ken, Japan, Fermentation ResearchInstitute Agency of Industrial Science and Technology, Ministry ofInternational Trade and Industry; Date of the Deposit: Oct. 5, 1984;Accession Number: FERM BP-927) or Streptomyces hygroscopicus Subsp.vakushimaensis No. 7238 (Depositary Authority: 1-3, Higashi 1 chome,Yatabe-machi, Tsukuba-gun, Ibaraki-ken, Japan, Fermentation ResearchInstitute Agency of Industrial Science and Technology, Ministry ofInternational Trade and Industry; Date of the Deposit: Jan. 12, 1985;Accession Number: FERM BP-928). Such situations are shown inEP-A-0184162.

Among those tricyclic compound (I), FK506 represented by the followingstructural formula is a typical compound.

Generic name: Tacrolimus

Chemical name:17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo-[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone.

On the other hand, a medicinal aerosol is a drug delivery system adaptedto deliver a medicinally active substance in a finely divided form alongwith inspired air into the recipient's airway for the treatment ofattacks of bronchial asthma, for instance, and is in broad use in thefield of medicine.

The conventional medicinal aerosol utilizes one or more kinds ofliquefied chlorofluorocarbons (hereinafter referred to collectively asCFC) as the propellant and is available in a system such that a finelydivided medicinally active substance has been dispersed in CFC with theaid of a suitable dispersant.

For example, aerosol of the tricyclic compound (I) comprising such CFCwere already suggested in WO 90/14826.

However, CFC is associated with the on-going destruction of the ozonelayer of the atmosphere and a worldwide total ban on its use is foreseenwithin this century. Under the circumstances, the use of liquefiedhydrofluoroalkanes (hereinafter sometimes referred to collectively asHFA) is being contemplated as substitute propellants for aerosols.However, despite their advantage of being lenient to the ozonosphere incomparison with CFC, HFA has the disadvantage that because of the totalinsolubility of the conventional dispersants (e.g. soybean lecithin)therein, medicinally active substances cannot be successfully dispersedin HFA.

To overcome the above disadvantage, an aerosol system providing for auniform dispersion of a medicinally active substance has been proposedwhich comprises HFA and, as a dispersant, a polymer containing anHFA-soluble amide or carboxylic ester as a recurrent unit (such aspolyvinylpyrrolidone, polyvinyl acetate, acrylic acid-methacrylic estercopolymer) (WO 93/05765).

The polymer used in the above aerosol system is a solid substance, withthe result that when a premix of the polymer with the active substanceis to be dispersed in the propellant, there occurs a segregation of theactive ingredient. Therefore, it is common practice to feed the activesubstance and the polymer respectively to a cooling agitation tank or apressure tank, then adding HFA under cooling or elevated pressure withstirring to disperse the active substance in the HFA, and distributingthe dispersion into dispensing containers. However, this procedure isnot only complicated but also has the disadvantage that because theproportion of the active ingredient is quite small, a uniformity of itscontent for each dispenser can hardly be insured in the stage ofportion-wise distribution of the propellant dispersing the activeingredient.

SUMMARY OF THE INVENTION

This inventors of this invention did much research for overcoming theabove-mentioned disadvantages and discovered that when a medium-chainfatty acid triglyceride is used as the dispersant in the manufacture ofa medicinal aerosol, the tricyclic compound (I) can be uniformlydispersed in HFA by kneading the tricyclic compound (I) with themedium-chain fatty acid triglyceride in the first place, distributingthe kneaded mass into aerosol dispensers, and filling the respectivedispensers with HFA under cooling or elevated pressure and that, as aresult, not only the aerosol preparation process is simplified but alsothe final aerosol has a minimal dispenser-to-dispenser variation incontent of the active ingredient. They accordingly have perfected thisinvention.

DETAILED DESCRIPTION OF THE INVENTION

The aerosol composition of this invention comprises a tricyclic compound(I) or a pharmaceutically acceptable salt thereof mentioned below, aliquefied hydrofluoroalkane, and a medium-chain fatty acid triglyceride.

The tricyclic compound (I) used in the present invention is representedby the following formula:

wherein each of adjacent pairs of R¹ and R², R³ and R⁴ or R⁵ and R⁶independently

(a) is two adjacent hydrogen atoms, or

(b) may form another bond formed between the carbon atoms to which theyare attached,

and further, R² may be an alkyl group;

R⁷ is a hydrogen atom, a hydroxy group, a protected hydroxy group or analkoxy group, or an oxo group together with R¹;

each of R⁸ and R⁹ is independently a hydrogen atom or a hydroxy group;

R¹⁰ is a hydrogen atom, an alkyl group, an alkyl group substituted byone or more hydroxy groups, an alkenyl group, an alkenyl groupsubstituted by one or more hydroxy groups or an alkyl group substitutedby an oxo group;

X is an oxo group, (a hydrogen atom and a hydroxy group), (a hydrogenatom and a hydrogen atom), or a group represented by the formula —CH₂O—;

Y is an oxo group, (a hydrogen atom and a hydroxy group), (a hydrogenatom and a hydrogen atom), or a group represented by the formulaN—NR¹¹R¹² or N—OR¹³;

each of R¹¹ and R¹² is independently a hydrogen atom, an alkyl group, anaryl group or a tosyl group;

each of R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²² and R²³ is independentlya hydrogen atom or an alkyl group;

each of R²⁰ and R²¹ is independently an oxo group or (R²⁰a and ahydrogen atom) or (R²¹a and a hydrogen atom) in which each of R²⁰a andR²¹a is independently a hydroxy group, an alkoxy group or a grouprepresented by the formula —OCH₂OCH₂CH₂OCH₃, or R²¹a is a protectedhydroxy group, or R²⁰a and R²¹a may together represent an oxygen atom inan epoxide ring;

n is an integer of 1, 2 or 3; and

in addition to the above definitions, Y, R¹⁰ and R²³, together with thecarbon atoms to which they are attached, may represent a saturated orunsaturated 5- or 6-membered nitrogen, sulfur and/or oxygen containingheterocyclic ring optionally substituted by one or more groups selectedfrom the group consisting of an alkyl, a hydroxy, an alkyl substitutedby one or more hydroxy groups, an alkoxy, a benzyl and a group of theformula —CH₂Se(C₆H₅).

Hereinafter, various terms which are included in the scope of thepresent invention will be defined:

Each definition in the formula (I) will be detailed as follows.

The term “lower” means, unless otherwise indicated, a group having 1 to6 carbon atoms. Preferable examples of the “alkyl groups” include astraight or branched chain aliphatic hydrocarbon residue, for example, alower alkyl group such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, pentyl, neopentyl and hexyl. Preferable examples of the“alkenyl groups” include a straight or branched chain aliphatichydrocarbon residue having one double-bond, for example, a lower alkenylgroup such as vinyl, propenyl (e.g., allyl group), butenyl,methylpropenyl, pentenyl and hexenyl. Preferable examples of the “arylgroups” include phenyl, tolyl, xylyl, cumenyl, mesityl and naphthyl.

Preferable protective groups in the “protected hydroxy groups” are1-(lower alkylthio) (lower)alkyl group such as a lower alkylthiomethylgroup (e.g., methylthiomethyl, ethylthiomethyl, propylthiomethyl,isopropylthiomethyl, butylthiomethyl, isobutylthiomethyl,hexylthiomethyl, etc.), more preferably C₁₋₄ alkylthiomethyl group, mostpreferably methylthiomethyl group; trisubstituted silyl group such as atri(lower)alkylsilyl (e.g., trimethylsilyl, triethylsilyl,tributylsilyl, tert-butyldimethylsilyl, tri-tert-butylsilyl, etc.) orlower alkyl-diarylsilyl (e.g., methyldiphenylsilyl, ethyldiphenylsilyl,propyldiphenylsilyl, tert-butyldiphenyl-silyl, etc.), more preferablytri(C₁₋₄)alkylsilyl group and C₁₋₄ alkyldiphenylsilyl group, mostpreferably tert-butyldimethylsilyl group and tert-butyldiphenylsilylgroup; or an acyl group such as an aliphatic, aromatic acyl group or analiphatic acyl group substituted by an aromatic group, which are derivedfrom a carboxylic acid, sulfonic acid or carbamic acid.

Examples of the aliphatic acyl groups include a lower alkanoyl groupoptionally having one or more suitable substituents such as carboxy,e.g., formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl,isovaleryl, pivaloyl, hexanoyl, carboxyacetyl, carboxypropionyl,carboxybutyryl, carboxyhexanoyl, etc.; acyclo(lower)alkoxy(lower)alkanoyl group optionally having one or moresuitable substituents such as lower alkyl, e.g., cyclopropyloxyacetyl,cyclobutyloxypropionyl, cycloheptyloxybutyryl, menthyloxyacetyl,menthyloxypropionyl, menthyloxybutyryl, menthyloxypentanoyl,menthyloxyhexanoyl, etc.; a camphorsulfonyl group; or a loweralkylcarbamoyl group having one or more suitable substituents such ascarboxy or protected carboxy, for example, carboxy(lower)alkylcarbamoylgroup (e.g., carboxymethylcarbamoyl, carboxyethylcarbamoyl,carboxypropylcarbamoyl, carboxybutylcarbamoyl, carboxypentylcarbamoyl,carboxyhexylcarbamoyl, etc.), tri-(lower)alkylsilyl(lower)alkoxycarbonyl (lower)alkylcarbamoyl group (e.g.,trimethylsilylmethoxycarbonylethylcarbamoyl,trimethylsilylethoxycarbonylpropylcarbamoyl,triethylsilylethoxycarbonylpropylcarbamoyl,tert-butyldimethylsilylethoxycarbonylpropylcarbamoyl,tri-methylsilylpropoxycarbonylbutylcarbamoyl, etc.) and so on.

Examples of the aromatic acyl groups include an aroyl group optionallyhaving one or more suitable substituents such as nitro, e.g., benzoyl,toluoyl, xyloyl, naphthoyl, nitrobenzoyl, dinitrobenzoyl,nitronaphthoyl, etc.; and an arenesulfonyl group optionally having oneor more suitable substituents such as halogen, e.g., benzenesulfonyl,toluenesulfonyl, xylenesulfonyl, naphthalenesulfonyl,fluorobenzenesulfonyl, chlorobenzenesulfonyl, bromobenzenesulfonyl,iodobenzenesulfonyl, etc.

Examples of the aliphatic acyl groups substituted by an aromatic groupinclude ar(lower)alkanoyl group optionally having one or more suitablesubstituents such as lower alkoxy or trihalo(lower)alkyl, e.g.,phenylacetyl, phenylpropionyl, phenylbutyryl,2-trifluoromethyl-2-methoxy-2-phenylacetyl,2-ethyl-2-trifluoromethyl-2-phenylacetyl,2-trifluoromethyl-2-propoxy-2-phenylacetyl, etc.

More preferable acyl groups among the aforesaid acyl groups are C₁₋₄alkanoyl group optionally having carboxy,cyclo(C₅₋₆)alkoxy(C₁₋₄)alkanoyl group having two (C₁₋₄)alkyls at thecycloalkyl moiety, camphorsulfonyl group, carboxy-(C₁₄)alkylcarbamoylgroup, tri(C₁₋₄)alkylsilyl(C₁₋₄)-alkoxycarbonyl(C₁₋₄)alkylcarbamoylgroup, benzoyl group optionally having one or two nitro groups,benzenesulfonyl group having halogen or phenyl(C₁₋₄)alkanoyl grouphaving C₁₋₄ alkoxy and trihalo(C₁₋₄)alkyl group. Among these, the mostpreferable ones are acetyl, carboxypropionyl, menthyloxyacetyl,camphorsulfonyl, benzoyl, nitrobenzoyl, dinitrobenzoyl,iodobenzenesulfonyl and 2-trifluoromethyl-2-methoxy-2-phenylacetyl.

Preferable examples of the “5- or 6-membered nitrogen, sulfur and/oroxygen containing heterocyclic ring” include a pyrrolyl group and atetrahydrofuryl group.

The pharmaceutically acceptable salt of the tricyclic compound (I)includes conventional non-toxic and pharmaceutically acceptable saltsuch as the salt with inorganic or organic bases, specifically, analkali metal salt such as sodium salt and potassium salt, an alkaliearth metal salt such as calcium salt and magnesium salt, an ammoniumsalt and an amine salt such as triethylamine salt andN-benzyl-N-methylamine salt.

With respect to the tricyclic compound (I), it is to be understood thatthere may be conformers and one or more stereoisomers such as opticaland geometrical isomers due to asymmetric carbon atom(s) and doublebond(s), and such conformers and isomers are also included within thescope of the present invention.

The tricyclic compound of the formula (I) and its salt can be in theform of a solvate, which is included within the scope of the presentinvention. The solvate preferably include a hydrate and an ethanolate.

FK506 is the most preferable compound belonging to the tricycliccompound (I). Other preferable compounds are listed hereinbelow.

1,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,17,21,27-pentamethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone,

12-[2-(4-acetoxy-3-methoxycyclohexyl)-1-methylvinyl]-17-allyl-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone,

17-allyl-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-12-[2-[4-(3,5-dinitrobenzoyloxy)-3-methoxycyclohexyl]-1-methylvinyl]-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]-octacos-18-ene-2,3,10,16-tetraone,

17-allyl-12-[2-[4-[(−)-2-trifluoromethyl-2-methoxy-2-phenylacetoxy]-3-methoxycyclohexyl]-1-methylvinyl]-1,14-dihydroxy-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)octacos-18-ene-2,3,10,16-tetraone.

17-ethyl-1,14-dihydroxy-12-[(2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone(FR900520), and

17-ethyl-1,14,20-trihydroxy-12-[2-(3,4-dihydroxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone.

The liquefied hydrofluoroalkane that can be used as the propellant inthe medicinal aerosol composition of this invention includes but is notlimited to 1,1,1,2-tetrafluoroethane (CH₂FCF₃, hereinafter HFA-134a) and1,1,1,2,3,3,3-heptafluoropropane (CF₃CHCF₃, hereinafter HFA-227) andthese liquefied hydrofluoroalkanes can be used either alone or incombination.

The medium-chain fatty acid triglyceride (MCT) for use as the dispersantfor the active ingredient in the aerosol composition of this inventionis predominantly composed of the triglycerides of saturated fatty acids[CH₃(CH₂)_(n)COOH, n=4-10], and such commercial products as Miglyol (thetrademark of Dynamit Novel) 812, Panacete (the trademark of NOFCorporation) 810, Coconard (the trademark of Kao Corporation), Myritol(the trademark of Hankel-Hakusui) GM, ODO (the trademark of The NisshinOil Mills, Ltd.), etc. can be utilized. The above MCT can be used eitheralone or in combination.

The formulating amount of said medium-chain fatty acid triglyceride isdependent on the type and quantity of the active ingredient but mayrange generally from 0.05 to 5 w/v % and preferably from 0.1 to 2 w/v %.

The above-mentioned tricyclic compound (I) or a pharmaceuticallyacceptable salt thereof used in the aerosol composition of the presentinvention is preferably in the form of fine particles. And in such case,it may be pulverized beforehand to a particle size of about 0.5-5 μm,more preferably 1-3 μm, by a conventional method, such as using jetmill. The amount of the tricyclic compound (I) or a pharmaceuticallyacceptable salt thereof contained in the present aerosol composition isthe therapeutically effective one, and varies from and depends on thetype of the aerosol composition and the age and condition of eachindimisual patient to be treated. However, it is generally 0.001-10 w/v% and preferably 0.005-5 w/v %.

Furthermore, the aerosol composition of this invention may furthercontain the conventional additives such as dispersant(s) (e.g.,polyvinylpyrrolidone, polyvinyl alcohol, sorbitan fatty acid ester,polyoxyethylene-sorbitan fatty acid ester (e.g. Tween 20, Span 85,etc.), fatty acid ester, polyethylene glycol-fatty acid ester,polyoxyethylene alkyl ether, sucrose ester, lecithin, HCO-60(polyoxyethylenehydrogenated castor oil), oleic acid, isopropylmyristate, etc.), in a proportion of 0.0001-0.05 w/v % and/orsolubilizer(s) for the tricyclic compound (I) or a pharmaceuticallyacceptable salt thereof (e.g., ethanol, glycerin, polyethylene glycol,propylene glycol, etc.), in a proportion of 1-20 w/v %.

The process for the preparation of the aerosol composition according tothis invention is characterized by kneading the tricyclic compound (I)or a pharmaceutically acceptable salt thereof and a medium-chain fattyacid triglyceride together, distributing the kneaded mass intodispensers, and filling the respective dispensers with a liquefiedhydrofluoroalkane under cooling or elevated pressure.

The more details of the process for preparation of the aerosolcomposition of the present invention are exemplified as follows.

First, the finely divided tricyclic compound (I) or a pharmaceuticallyacceptable salt thereof is kneaded with said medium-chain fatty acidtriglyceride and optional additives, such as polyvinylpyrrolidone or thelike, and the kneaded mass is distributed into dispensing containers(usually aluminum cans). Then, each resulting dispenser is filled withthe liquefied hydrofluoroalkane precooled to −20° C. to disperse theactive ingredient in the hydrofluoroalkane. The dispenser is then fittedwith a valve to provide a finished product.

As an alternative, after distributing the above kneaded mass intodispensing containers, each resulting dispenser may be fitted with avalve and, then, filled with said liquefied hydrofluoroalkane under anelevated pressure of 20-30 atmospheres at ordinary temperature.

The ejection amount of the medicinal aerosol of this invention is 25-150μl per valve actuation. Depending on the amount of the active substance,1-3 valve actuations are made per dose and 1-5 doses are administered aday.

EFFECT OF THE INVENTION

(1) The tricyclic compound (I) or its salt is insoluble or indispersiblein liquefied hydrofluoroalkanes, even if conventional dispersants, suchas soya lecithin, are admixed with.

However, by the addition of medium-chain fatty acid triglyceride (MCT),rot. only the improvement of dispersing condition of the tricycliccompound (I) but also the dramatic enhancement of the solubility of thetricyclic compound (I) in liquefied hydrofluoroalkanes were achieved.

As shown in Table 1, the solubility of FK506, which was used as arepresentative of the tricyclic compound (I), was increased up by mixingMCT into liquefied hydrofluoroalkanes. The addition of MCT enables thefilling of FK506 as a solution into aerosol system. As a result, thechange of spray performance will not be caused by aggregation of FK506crystalline particles and the emitted dose uniformity of FK506 can bemore reliable. The aerosol compositions used in this study were preparedaccording to a similar manner to that of Example 2.

TABLE 1 Effect of MCT Content on the Solubility of FK506 in HFAs MCTFK506 content (w/v %) FK506 content (w/v %) content in HFA-227 inHFA-134a (%) 0.05 0.1 0.2 0.5 0.05 0.2 0.05 ∘ — — — ∘ — 0.5 ∘ ∘ — — ∘ —2 ∘ ∘ ∘ — ∘ ∘ 5 ∘ ∘ ∘ — ∘ ∘ ∘: solution —: suspension

Moreover, since the medium-chain fatty acid triglyceride has an oilyconsistency at room temperature, it car be well 35 kneaded with thetricyclic compound (I) and after distributing the resulting kneaded massinto the dispensers, HFA can be filled thereinto under cooling orelevated pressure. The above achieved a remarkable uniformity of thecontent of the tricyclic compound (I) per dispenser.

Therefore, there is no variation in the delivery dose of the activeingredient on valve actuation.

The form of the aerosol composition of the present invention can be asolution-type or a suspension-type.

Therefore, depending on the amount of the content of the tricycliccompound (I) or its pharmaceutically acceptable salt thereof and/or MCT,the form of the aerosol composition of the present invention can beselective.

(2) Further, the addition of MCT was found to generate the novelcharacteristics in FK506 aerosol composition. For instance, the massmedian aerodynamic diameter (MMAD) calculated from the aerodynamicparticle size distribution as mentioned below, increased in proportionto MCT amount added (Table 2).

Aerodynamic Particle Size Distribution

According to the conventional method in USP23 (Apparatus 1), theaerodynamic particle size distribution was assessed from FK506 amount ineach stage after applying one mg FK506 to multistage cascade impactor byfiring FK506 aerosol composition. FK506 measurement was conducted byHPLC method and the MMAD was calculated from the particle sizedistribution. The FK506 aerosol compositions were prepared according toa similar manner to that of the below-mentioned Example 2.

TABLE 2 Effect of MCT Content on Aerodynamic Particle Size of 0.05%FK506 Aerosol Composition with HFAs. MCT content Mass Median AerodynamicDiameter Propellant (%) (μm) HFA-227 0.05 1.5 0.5 1.7 1 2.5 2 3.1 5 4.0HFA-134a 0.5 1.6

(3) Moreover, FK506 release rate from the mist particles was studiedaccording to the below-mentioned Dissolution Test. Thereby, it wasconfirmed that the release rate of FK506 was declined with addition ofMCT as presented in Table 3. Especially, such a release rate was apt tobe slower in solution than in suspension. These results clarified thatFK506 release rate can be regulated by controlling the amount of MCT.

Dissolution Test

FK506 dissolution from the mist particles after firing FK506 aerosolcomposition was examined in distilled water at 37° C. using the paddlemethod at 50 rpm, according to the dissolution test method in JP12. Theemitted dose from aerosol composition was adjusted to be one mg FK506 asa total in the test fluid. FK506 was measured by HPLC method. The FK506aerosol compositions were prepared according to a similar manner to thatof Example 2.

TABLE 3 Effect of MCT Content on Dissolution Rate of FK506 MCT content T50% (min) Propellant (%) FK506 0.05% FK506 0.2% HFA-227 0 5 9 0.5 30 121 38 15 2 43 28 5 51 37 HFA-134a 0.5 29 11 2 41 25

These novel characters suggested to enable to optimize the selectivityof pulmonary drug delivery and adjust drug absorption rate at deliveredsite, which means that the tricyclic compound (I) or a pharmaceuticallyacceptable salt thereof can be released sustainedly and that itstoxicity can be reduced thereby.

Industrial Field of Utilization

The aerosol composition of the present invention is useful for thetreatment and/or prevention of various diseases topically and/orsystemically.

Especially, due to the pharmacological activities of the tricycliccompound (I), the aerosol composition comprising it of the presentinvention is useful for the treatment and/or prevention of reversibleobstructive airways disease, which includes conditions such as asthma(e.g. bronchial asthma, allergic asthma, intrinsic asthma, extrinsicasthma and dust asthma), particularly chronic or inveterate asthma (e.g.late asthma and airway hyper-responsiveness), bronchitis and the like.

And further, the aerosol composition according to the present invention,due to the pharmacological activities, such as immunosuppressiveactivity and antimicrobial activity, of the tricyclic compound (I), isuseful for the treatment and/or prevention of immune-mediated diseasessuch as rejection by transplantation of organs or tissues such as heart,kidney, liver, bone marrow, skin, cornea, lung, pancreas, smallintestine, limb, muscle, nerve, intervertebral disk, trachea, etc.;graft-versus-host diseases by medulla ossium transplantation; autoimmunediseases such as rheumatoid arthritis, systemic lupus erythematosus,Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type Idiabetes, and the like; and further infectious diseases caused bypathogenic microorganisms.

Further, the aerosol composition of the present invention is also usefulfor the treatment and the prophylaxis of inflammatory andhyperproliferative skin diseases and cutaneous manifestations ofimmunologically-mediated illnesses, such as psoriasis, atopicdermatitis, contact dermatitis, eczematous dermatitis, seborrhoeicdermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysisbullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneouseosinophilias, lupus erythematosus, acne and alopecia areata;

various eye diseases such as autoimmune diseases and so on (e.g.keratoconjunctivitis, vernal conjunctivitis, uveitis associated withBehcet's disease, keratitis, herpetic keratitis, conical keratitis,dystrophia epithelialis corneae, corneal leukoma, ocular pemphigus,Mooren's ulcer, scleritis, Graves' ophthalmopathy, Vogt-Koyanagi-Haradasyndrome, sarcoidosis, etc.);

inflammation of mucosa and blood vessels such as gastric ulcers,vascular injury caused by ischemic diseases and thrombosis, ischemicbowel disease, enteritis, necrotizing enterocolitis, intestinal lesionsassociated with thermal burns, leukotriene B₄-mediated diseases;

intestinal inflammations/allergies such as coeliac disease, proctitis,eosinophilic gastroenteritis, mastocytosis, Crohn's disease andulcerative colitis;

food related allergic diseases which have symptomatic manifestationremote from the gastro-intestinal tract, for example, migraine, rhinitisand eczema;

renal diseases such as interstitial nephritis, Good-pasture's syndrome,hemolytic-uremic syndrome and diabetic nephropathy;

nervous diseases such as multiple myositis, Guillain-Barré syndrome,Ménière's disease and radiculopathy;

endocrine diseases such as hyperthyroidism and Basedow's disease;

hematic diseases such as pure red cell aplasia, aplastic anemia,hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmunehemolytic anemia, agranulocytosis, pernicious anemia, megaloblasticanemia and anerythroplasia;

bone diseases such as osteoporosis;

respiratory diseases such as sarcoidosis, pulmonary fibrosis andidiopathic interstitial pneumonia; skin diseases such asdermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photoallergicsensitivity and cutaneous T cell lymphoma;

circulatory diseases such as arteroiosclerosis, atherosclerosis,aortitis syndrome, polyarteritis nodosa and myocardosis;

collagen diseases such as scleroderma, Wegener's granuloma and Sjögren'ssyndrome;

adiposis;

eosinophilic fasciitis;

periodontal disease such as lesion of gingiva, periodontium, alveolarbone, substantia ossea dentis;

nephrotic syndrome such as glomerulonephritis;

male pattern alopecia or alopecia senilis;

muscular dystrophy;

pyoderma and Sezary's syndrome;

Addison disease;

active oxygen-mediated diseases, for example, organ injury such asischemia-reperfusion injury of organs (e.g. heart, liver, kidney,digestive tract) which occurs on preservation, transplantation orischemic diseases (e.g. thrombosis, cardiac infarction): intestinaldiseases such as endotoxin-shock, oseudomembranous colitis, colitiscaused by drug or radiation: renal diseases such as ischemic acute renalinsufficiency, chronic renal insufficiency: pulmonary diseases such astoxicosis caused by lung-oxygen or drug (e.g. paracort, bleomycins),lung cancer, pulmonary emphysema: ocular diseases such as cataracta,siderosis, retinitis, pigmentosa, senile macular degeneration, vitreousscarring, corneal alkali burn: dermatitis such as erythema multiforme,linear IgA bullous dermatitis, cement dermatitis: and others such asgingivitis, periodontitis, sepsis, pancreatitis, diseases caused byenvironmental pollution (e.g. air pollution), aging, carcinogens,metastasis of carcinoma, hypobaropathy;

diseases caused by histamine or leukotrience C₄ release;

and so on.

And further, the tricyclic compound (I) has liver regenerating activityand/or activities of stimulating hypertrophy and hyperplasia ofhepatocytes. Therefore, the present aerosol composition is useful forthe treatment and/or prevention of hepatic diseases such as immunogenicdiseases (e.g. chronic autoimmune liver diseases such as the groupconsisting of autoimmune hepatic disease, primary biliary cirrhosis andsclerosing cholangitis), partial liver resection, acute liver necrosis(e.g. necrosis caused by toxins, viral hepatitis, shock or anoxia),hepatitis B, hepatitis non-A/non-B, cirrhosis and hepatic failure suchas fulminant hepatitis late-onset hepatitis and “acute-on-chronic” liverfailure (acute liver failure on chronic liver diseases).

And further, the present aerosol composition is useful for variousdiseases because of its useful pharmacological activity such asaugmenting activity of chemotherapeutic effect, preventing or treatingactivity of cytomegalovirus infection, anti-inflammatory activity, andso on.

The aerosol composition of the present invention can also be obtainedwhen the compounds disclosed in patent applications such as EP-A-353678,Japanese Patent Application No. 2(1990)-74330, PCT/GB90/01262,EP-A-413532, PCT/JP91/00314, British Patent Applications No. 9012963.6,No. 9014136.7, No. 9014681.2, No. 9014880.0, No. 9014881.8, No.9015098.8, No. 9016115.9, and No. 9016693.5, EP-A-323865, EP-A-349061,EP-A-358508, EP-A-364031, EP-A-364032, EP-A-378317, EP-A-378320,EP-A-378321, EP-A-388153, EP-A-396399, EP-A-396400, EP-A-399579,EP-A-403242, EP-A-428365, EP-A-356399, GB 2225576 A, EP-A-402931,EP-A-427680, EP-A-445975, EP-A-455427, EP-A-463690, EP-A-464895,EP-A-466365, EP-A-478235, EP-A-480623, EP-A-509753, EP-A-515071,EP-A-520554, EP-A-526934, EP-A-530888, EP-A-532089, and EP-A-532088,W092/06992, WO92/20688, WO93/04679, WO93/05059, and WO93/04680, U.S.Pat. No. 5,149,701, German Patent Applications A-4021404, A-4028664,A-4028665, A-4028666, A-4028667, A-4028675, A-4028676, A-4028677,A-4028678, and A-4039587; and rapamycins such as rapamycin are employedinstead of the tricyclic compound (I) or its pharmaceutically acceptablesalt.

The present invention will be described hereinbelow with reference tothe following Examples, but it is not intended to limit the scope of theinvention.

EXAMPLE 1

FK506 was finely divided to a particle size of 2-3 μm by using a jetmill and the resulting powders were kneaded with Miglyol 812.

After distribution of the kneaded mass, each dispenser was filled withHFA-227 cooled to −20° C. beforehand and fitted with a valve to providean aerosol product containing the following ingredients per unit (5 ml).(cold filling method)

FK506 10 mg (0.2 (w/v)%) Miglyol 812 25 mg (0.5 (w/v)%) HFA-227 5 ml

EXAMPLE 2

Dispensers were charged with the kneaded mass containing the followingingredients per unit (5 ml) which were obtained according to a similarmanner to that of Example 1 and, after installation of the valve, eachdispenser was filled with HFA-227 pressurized to 20 atms at roomtemperature to provide a medicinal aerosol composition of the samecomposition as that of Example 1. (Pressure filling method)

FK506 5 mg (0.1 (w/v)%) Miglyol 812 10 mg (0.2 (w/v)%) HFA-227 5 ml

EXAMPLES 3-11

The following aerosol compositions were provided in the same manner asExample 1 or Example 2.

Tricyclic compound Medium-chain fatty (Content acid triglyceridePropellant Examples (w/v%)) (w/v%) (5 ml) 3 FK506 Miglyol 812 HFA-227 (0.05)  (0.05) 4 FK506 Miglyol 812 HFA-227 (0.1) (0.5) 5 FK506 Miglyol812 HFA-227 (0.2) (2)   6 FK506 Miglyol 812 HFA-227 (0.5) (5)   7 FK506Miglyol 812 HFA-134a  (0.05)  (0.05) 8 FK506 Miglyol 812 HFA-134a (0.2)(5)   9 FK506 Panacete 810 HFA-134a (0.1) (0.2) 10 FK506 CoconardHFA-134a (0.4) (1)   11 FR900520 Miglyol 812 HFA-227 (0.1) (0.2)

EXAMPLE 12

The aerosol composition containing the following ingredients per unit (5ml) was also prepared according to a similar manner to that of Example2.

FK506 10 mg Miglyol 812 25 mg Polyvinylpyrrolidone 0.25 mg HFA-227 5 ml

What is claimed is:
 1. An aerosol composition comprising a tricycliccompound (I) of the following formula:

wherein each of adjacent pairs of R¹ and R², R³ and R⁴ or R⁵ and R⁶ isindependently (a) two adjacent hydrogen atoms, or (b) may form anotherbond formed between the carbon atoms to which they are attached, andfurther, R² may be an alkyl group; R⁷ is a hydrogen atom, a hydroxygroup, a protected hydroxy group, or an alkoxy group or an oxo grouptogether with R¹; each of R⁸ and R⁹ is independently a hydrogen atom ora hydroxy group; R¹⁰ is a hydrogen atom, an alkyl group, an alkyl groupsubstituted by one or more hydroxy groups, an alkenyl group, an alkenylgroup substituted by one or more hydroxy groups or an alkyl groupsubstituted by an oxo group; X is an oxo group, (a hydrogen atom and ahydroxy group), (a hydrogen atom and a hydrogen atom), or a grouprepresented by the formula —CH₂O—; Y is an oxo group, (a hydrogen atomand a hydroxy group), (a hydrogen atom and a hydrogen atom), or a grouprepresented by the formula N—NR¹¹R¹² or N—OR¹³; each of R¹¹ and R¹² isindependently a hydrogen atom, an alkyl group, an aryl group or a tosylgroup; each of R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²² and R²³ isindependently a hydrogen atom or an alkyl group; each of R²⁰ and R²¹ isindependently an oxo group, (R²⁰a and a hydrogen atom) or (R²¹a and ahydrogen atom), in which each of R²⁰a and R²¹a is independently ahydroxy group, an alkoxy group or a group represented by the formula—OCH₂OCH₂CH₂OCH₃, or R²¹a is a protected hydroxy group, or R²⁰a and R²¹amay together represent an oxygen atom in an epoxide ring; n is aninteger of 1 or 2; and in addition to the above definitions, Y, R¹⁰ andR²³ may represent a pyrrolyl ring or a tetrahydrofuryl ring optionallysubstituted by one or more groups selected from the group consisting ofan alkyl, a hydroxy, an alkyl substituted by one or more hydroxy groups,an alkoxy, a benzyl and a group of the formula —CH₂Se(C₆H₅); or apharmaceutically acceptable salt thereof, a liquefied hydrofluoroalkaneand a medium-chain fatty acid triglyceride.
 2. The aerosol compositionas claimed in claim 1, in which the tricyclic compound (I) or apharmaceutically acceptable salt thereof is contained in amount of 0.001to 10 w/v %.
 3. The aerosol composition as claimed in claim 1, in whichthe tricyclic compound (I) is the one therein each of adjacent pairs ofR³ and R⁴ or R⁵ and R⁶ independently may form another bond formedbetween the carbon atoms to which they are attached; each of R⁸ and R²³is independently a hydrogen atom; R⁹ is a hydroxy group; R¹⁰ is a methylgroup, an ethyl group, a propyl group or an allyl group; X is (ahydrogen atom and a hydrogen atom) or an oxo group; Y is an oxo group;each of R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹ and R²² is a methyl group; each ofR²⁰ and R²¹ is independently (R²⁰a and a hydrogen atom) or (R²¹a and ahydrogen atom) in which each of R²⁰a and R²¹a is a hydroxy group or analkoxy group, or R²¹a is a protected hydroxy group; and n is an integerof 1 or
 2. 4. The aerosol composition as claimed in claim 3, in whichthe tricyclic compound (I) is the one wherein R⁷ is a hydrogen atom, ahydroxy group or a protected hydroxy group; X is an oxo group; R²⁰a is amethoxy group; R²¹a is a hydroxy group or a protected hydroxy group. 5.The aerosol composition as claimed in claim 4, in which the tricycliccompound (I) is17-allyl-1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0^(4,9)]octacos-18-ene-2,3,10,16-tetraone.6. The aerosol composition as claimed in claim 1, in which the liquefiedhydrofluoroalkane is 1,1,1,2-tetrafluoroethane.
 7. The aerosolcomposition as claimed in claim 1, in which the medium-chain fatty acidtriglyceride is CH₃(CH₂)_(n)COOH, wherein n is 4-10.
 8. The aerosolcomposition as claimed in claim 1, which further comprises an optionaladditive selected from the group consisting of polyvinylpyrrolidone andethanol.
 9. A process for a preparation of the aerosol composition asclaimed in claim 1, comprising; (1) kneading said tricyclic compound (I)or a pharmaceutically acceptable salt thereof with a medium-chain fattyacid triglyceride, (2) distributing the resulting kneaded mass intodispensers, and (3) filling each dispenser with said liquefiedhydrofluoroalkane under cooling or elevated pressure.
 10. The aerosolcomposition as claimed in claim 1, in which the liquefiedhydrofluoroalkane is 1,1,1,2,3,3,3-heptafluoropropane.
 11. The aerosolcomposition as claimed in claim 1, in which said medium-chain fatty acidtriglyceride is present in an amount range of from 0.05 to 5 w/v %. 12.The aerosol composition as claimed in claim 1, in which saidmedium-chain fatty acid triglyceride is present in an amount range offrom 0.1 to 2 w/v %.
 13. The process as claimed in claim 9, wherein saidmedium-chain fatty acid triglyceride is CH₃(CH₂)_(n)COOH, wherein n is4-10.